Han T L, Ford J H, Webb G C, Flaherty S P, Correll A, Matthews C D
Genetics Department, University of Adelaide, Queen Elizabeth Hospital, Woodville, South Australia.
Mol Reprod Dev. 1993 Mar;34(3):308-13. doi: 10.1002/mrd.1080340311.
Double fluorescence in situ hybridization (FISH) was used to detect sex chromosomes in decondensed human sperm nuclei. Biotinylated X chromosome specific (TRX) and digoxigenin-labeled Y chromosome specific (HRY) probes were simultaneously hybridized to sperm preparations from 12 normal healthy donors. After the hybridization, the probes were detected immunocytochemically, using two different and independent affinity systems. Ninety-six percent of the 12,636 sperm showed fluorescent labeling, of which 47.4% were haploid X and 46.8% were haploid Y. A frequency of 0.46% of XX-bearing sperm (0.28% disomic, 0.18% diploid) and 0.38% YY-bearing sperm (0.21% disomic, 0.17% diploid) was found. The overall proportions of X- and Y-bearing sperm in the ejaculates were 47.9% and 47.2%, respectively, which was not significantly different from the expected 50:50 ratio. In addition 0.21% of cells appeared to be haploid XY-bearing sperm, 0.62% were diploid XY-bearing cells, and 0.05% of cells were considered to be tetraploid cells. The application of double FISH to human sperm using X-chromosome and Y-chromosome probes has allowed a more accurate assessment of the sex chromosal complements in sperm than single FISH method and quinacrine staining for Y-bodies.
采用双重荧光原位杂交（FISH）技术检测去浓缩的人类精子细胞核中的性染色体。将生物素化的X染色体特异性（TRX）探针和地高辛配体标记的Y染色体特异性（HRY）探针同时与12名正常健康供者的精子样本进行杂交。杂交后，使用两种不同且独立的亲和系统通过免疫细胞化学方法检测探针。在12636个精子中，96%显示出荧光标记，其中47.4%为单倍体X精子，46.8%为单倍体Y精子。发现含XX精子的频率为0.46%（双体0.28%，二倍体0.18%），含YY精子的频率为0.38%（双体0.21%，二倍体0.17%）。射精中含X和含Y精子的总体比例分别为47.9%和47.2%，与预期的50:50比例无显著差异。此外，0.21%的细胞似乎是含单倍体XY的精子，0.62%是含二倍体XY的细胞，0.05%的细胞被认为是四倍体细胞。与单FISH方法和Y小体的喹吖因染色相比，使用X染色体和Y染色体探针的双重FISH技术可更准确地评估精子中的性染色体组成。
